shRNA gene silencing Human Neuroblastoma cells (SH-SY5Y) Connexin 43

Get tips on using QIAamp DNA FFPE Tissue Kit to perform DNA isolation / purification Cells - Primary cells Pseudomyxoma peritonei (PMP) cells

Get tips on using Wizard® Genomic DNA Purification Kit to perform DNA isolation / purification Cells - Primary cells Rat cortical neurons

Get tips on using EMD Millipore™ Chemicon™ CpGenome™ Universal DNA Modification Kit to perform DNA methylation profiling Gene specific profiling - HepG2 FHIT

Get tips on using EMD Millipore™ Chemicon™ CpGenome™ Universal DNA Modification Kit to perform DNA methylation profiling Gene specific profiling - Hep3B SFRP3

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat mesenchymal stem cells (rMSC)

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat schwann cells

Get tips on using peSpCas9(1.1)-2×sgRNA (empty, donor) to perform CRISPR Human - Repression SLC7A11

Get tips on using CDX2 to perform Immunohistochemistry Human - CDX2

Plasmid isolation is an important technique in molecular biology or any kind of genetic editing. It involves amplifying plasmids overnight by transforming them into competent bacterial cells. The desired colonies of these bacteria can then be grown in shaker cultures, at appropriate shaking speed, oxygen availability and temperature. These liquid cultures can then be ultracentrifuged to pellet the bacteria, which are then used for plasmid isolation. The bacteria are first resuspended in a buffer, then lysed, neutralized, purified in a column, eluted, precipitated with ethanol and then resuspended. During plasmid isolation, it is important to lyse cells quickly because lysing bacteria for too long may lead to irreversible denaturing of the plasmid. Usually, alkaline lysis is used for isolation because it is a mild treatment. It isolates plasmid DNA and other cell components such as proteins by breaking cells apart with an alkaline solution. Precipitation removes the proteins, and the plasmid DNA recovers with alcohol precipitation. Resuspension and lysis buffers should be mixed thoroughly in order to prevent the DNA from breaking into smaller fragments. This is because broken gDNA can reanneal and remain in the solution, without binding to the column.

Get tips on using Wizard® Genomic DNA Purification Kit to perform DNA isolation / purification Cells - Primary cells Mouse embryonic fibroblast (MEF)